Cardiolipin binding a light chain from lupus-prone mice.
Autoantibodies in systemic lupus erythematosus react with multiple epitopes on highly conserved molecules such as nucleic acids, cytoskeletal proteins, phospholipids, and phospholipid-binding proteins. Analysis of the heavy- and light-chain variable sequences (VH and VL) has shown that a restricted set of V genes gives rise to these autoantibodies. Several monoclonal antibodies were developed from a strain of mouse prone to lupus (F1 male NZW x BXSB). Two of these antibodies, A1.72 and A1.84, reacted directly with cardiolipin and their VH and VL sequences were analyzed. Surprisingly, these two antibodies had identical light-chain variable sequences despite having substantially different heavy-chain variable sequences. This VL sequence, VL 72/84 was 97% identical with the germ-line sequences with only four single nucleotide substitutions. When this VL sequence was shuffled with the VH sequence of other monoclonal antibodies and expressed as single chain variable fragment (scFv) in Escherichia coli, it imparted cardiolipin-binding activity to the hybrids. Furthermore, the VL 72/84 sequence, when expressed alone without any VH sequence, also bound to cardiolipin. The antibodies and their recombinant fragments were immunoaffinity-purified on cardiolipin liposomes. The dissociation constant of the light chain for cardiolipin was similar to the intact molecule (21 +/- 0.01 vs 20 +/- 0.03 nM). These studies demonstrate that the VL sequence alone, in the absence of any other immunoglobulin domains, can mediate cardiolipin binding, raising the possibility that antigen specificity of certain antibodies may exclusively reside in their light-chain sequences.
Pereira
B
B
Department of Internal Medicine, University of Texas Health Science Center, Houston 77030, USA.
Benedict
C R
CR
Le
A
A
Shapiro
S S
SS
Thiagarajan
P
P
D000595:Amino Acid Sequence; D000818:Animals; D017153:Antibodies, Anticardiolipin / Q000378:metabolism / Q000502:physiology; D000911:Antibodies, Monoclonal / Q000378:metabolism / Q000502:physiology; D000915:Antibody Affinity; D000918:Antibody Specificity; D001483:Base Sequence; D001666:Binding Sites, Antibody; D002308:Cardiolipins / Q000276:immunology* / Q000378:metabolism*; D002417:Cattle; D003433:Crosses, Genetic; D004198:Disease Susceptibility; D007147:Immunoglobulin Light Chains / Q000378:metabolism* / Q000502:physiology; D008180:Lupus Erythematosus, Systemic / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D008297:Male; D051379:Mice; D008814:Mice, Inbred NZB; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000276:immunology
D017153:Antibodies, Anticardiolipin; D000911:Antibodies, Monoclonal; D002308:Cardiolipins; D007147:Immunoglobulin Light Chains